Bacillus amyloliquefaciens B011 for preventing and treating tobacco bacterial wilt and application thereof
A technology for resolving amyloliquefaciens and tobacco bacterial wilt, which is applied to Bacillus amyloliquefaciens B011 and its application fields, can solve the problems such as the lack of specific medicines for preventing and controlling tobacco bacterial wilt, the environment of bacterial resistance, and the impact on the control effect, etc. Good development prospects and application potential, no toxic side effects, good control effects
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Embodiment 1
[0045] The separation method of Bacillus amyloliquefaciens B011 of the present embodiment, its separation steps are as follows:
[0046] (1) Sampling: in the tobacco field where tobacco bacterial wilt has occurred, select still growing healthy tobacco plant samples, wash, cut off the main root and lateral roots and carry out surface disinfection, get a sterilized sample, and set aside;
[0047] (2) Grinding: put 0.1 g of the spare sterilized sample in a sterilized mortar, add 10 ml of sterile water for grinding to obtain a grinding liquid, and set aside;
[0048] (3) Centrifugation: Take 1ml of the spare grinding solution and let it stand for 30 minutes, and then centrifuge it with a centrifuge at 7000r / min at 4°C to obtain the supernatant after centrifugation, and set it aside;
[0049] (iv) Purification: use sterile water for 10 minutes to reserve the supernatant -5 After gradient dilution, use the coating plate method on the NA medium, and after the colonies grow on the se...
Embodiment 3
[0055] The bacillus amyloliquefaciens B011 of application embodiment one gained prepares bacillus amyloliquefaciens B011 bacteriostatic agent:
[0056] First, prepare the NA medium slant, inoculate the Bacillus amyloliquefaciens B011 obtained in Example 1, put it into a 30°C incubator and cultivate it for 48 hours to obtain the slant bacteria, and set aside;
[0057] Next, configure NA liquid medium, divide into triangular flasks (250mL, 100mL liquid) before sterilization, sterilize at 121°C for 20min, and wash off the above-mentioned slanted bacteria directly with sterile water on the ultra-clean workbench after cooling. Insert it into the Erlenmeyer flask containing the above-mentioned sterilized NA liquid medium, culture in a shake flask, pH 7.0, 29°C, 170r / min, and cultivate for 48h to obtain shake flask culture bacteria for future use;
[0058] Next, configure the NA liquid medium according to the optimum fermentation condition combination formula optimized by the orthogo...
Embodiment 5
[0065] The Bacillus amyloliquefaciens B011 bacteriostatic agent of application embodiment three gained is to the inhibitory action test of multiple plant pathogenic bacteria:
[0066] Adopt plate confrontation method, test application embodiment three gained Bacillus amyloliquefaciens B011 bacteriostatic agent to tobacco black shank bacterium (P.parasitica), tobacco red spot bacterium (A.alternata Keissler), capsicum anthracnose bacterium (C.capsici), The antagonistic effects of seven pathogenic fungi, including P.capsici, C.gloeosprioides, P. oryzae and C.cuc-umerinum, were tested in PDA The central access of the tablet is 5mm 2 The target mycelium block of the target hyphae block is punched according to the method described in "Example 2" at the same distance from the target mycelium block. Add 20 μL of the Bacillus amyloliquefaciens B011 bacteriostatic agent prepared according to "Example 3" to two symmetrical wells, and inject sterile NA liquid medium into the other two s...
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